Systems and methods for preparing a thrombin serum

ABSTRACT

A system for preparing a thrombin serum that can include a containment device, a cage received within the containment device, a cap attachable to the containment device, an inlet port configured to introduce a non-anti-coagulated autologous blood fluid into the containment device, and an outlet port. An activator, such as glass beads, can be present within the containment device.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/812,592, filed Mar. 9, 2020, now U.S. Pat. No. 11,045,526, which is adivisional of U.S. application Ser. No. 15/455,417, filed on Mar. 10,2017, now U.S. Pat. No. 10,596,236, issued Mar. 24, 2020, which claimsthe benefit of U.S. Provisional Application No. 62/306,304, filed onMar. 10, 2016, the entirety of which are incorporated herein byreference.

BACKGROUND

This disclosure relates to systems and methods for preparing a thrombinserum.

Thrombin is an enzyme in blood plasma that clots blood by convertingfibrinogen to fibrin. Thrombin has many surgical uses and can be used totreat wounds or control bleeding during surgical procedures.

SUMMARY

This disclosure describes systems and methods for preparing a thrombinserum. The thrombin serum can be used to produce a clotted product.

An exemplary system for preparing a thrombin serum includes acontainment device and an activator (e.g., a plurality of beads orspheres) to artificially start the coagulation cascade after adding anautologous blood fluid to the containment device. The system can producea thrombin serum from a non-anticoagulated autologous blood fluid. Onceprepared, the thrombin serum can be added to platelet rich plasma orother autologous blood fluids to produce a clot.

A system for preparing a thrombin serum according to an exemplary aspectof the present disclosure includes, inter alia, a containment device, acage received within the containment device, a cap attached to thecontainment device, an inlet port configured to introduce anon-anti-coagulated autologous blood fluid into the containment device,and an outlet port configured to remove a thrombin serum from thecontainment device.

A method for preparing a thrombin serum according to another exemplaryaspect of the present disclosure includes, inter alia, adding a firstamount of a non-anti-coagulated autologous blood fluid to a containmentdevice, incubating the containment device, extracting a thrombin serumfrom containment device, and adding the thrombin serum to a secondautologous blood fluid to produce a clotted product.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a system for preparing a thrombin serum.

FIG. 2 illustrates a cross-sectional view of the system of FIG. 1 .

FIG. 3 illustrates a cage of the system of FIG. 1 .

FIG. 4 illustrates a filter of the system of FIG. 1 .

FIGS. 5A and 5B illustrate a tray assembly for packaging a system forpreparing a thrombin serum.

FIG. 6 schematically illustrates harvesting an autologous blood fluidsample from a patient.

FIG. 7 illustrates adding an autologous blood fluid to a containmentdevice of a system for preparing a thrombin serum.

FIG. 8 schematically illustrates incubating the containment device.

FIG. 9 schematically illustrates adding additional autologous bloodfluid to the containment device.

FIG. 10 schematically illustrates additional steps for preparing athrombin serum within the containment device.

FIG. 11 schematically illustrates withdrawing a thrombin serum from thecontainment device.

FIGS. 12A and 12B schematically illustrate adding a thrombin serum to asecond autologous blood fluid to produce a clot.

FIGS. 13A and 13B schematically illustrate exemplary surgical uses of aclotted product that is produced by adding a thrombin serum to anautologous blood fluid.

DETAILED DESCRIPTION

This disclosure describes systems and methods for preparing a thrombinserum. Once prepared, thrombin serum can be used to produce a clot in anautologous blood fluid such as platelet rich plasma (PRP). The clottedproduct is then used at the point of care to treat a patient.

In some embodiments, a system for preparing the thrombin serum includesa containment device and an activator, such as a plurality of beads,received within the containment device. An activator such as glass beadscan artificially start the coagulation cascade when autologous bloodfluid is added to the containment device. A thrombin serum can beproduced via the methods described herein, which include contacting anactivator with an autologous blood fluid (e.g., whole blood,platelet-rich plasma (PRP), platelet-poor plasma (PPP), bone marrowaspirate (BMA), bone marrow concentrate (BMC), or combinations thereof).The thrombin serum may be extracted from the containment device and thenadded to a second autologous blood fluid to produce a clot. These andother features are discussed in greater detail in the followingparagraphs of this detailed description.

FIGS. 1 and 2 illustrate a system 10 for preparing a thrombin serum.Thrombin is an enzyme in blood plasma that clots blood by convertingfibrinogen to fibrin. A thrombin serum is a blood serum comprisingthrombin at greater than basal levels. The thrombin serums of thisdisclosure may therefore be used in a wide variety of surgicalprocedures, including but not limited to controlling bleeding, treatingwounds, augmenting tissue repairs, repairing/plugging voids in tissue orbone, etc.

In an embodiment, a method of preparing a thrombin serum includescontacting an autologous blood fluid with an activator. Followingcontact, the autologous blood fluid is incubated with the activator in acontainment device to produce a thrombin serum. The incubation may occurat room temperature. The autologous blood fluid may include whole blood,PRP, PPP, BMA, BMC, or combinations thereof, for example. In anembodiment, an autologous blood fluid contacts and is incubated with anactivator without any other substance present. In an embodiment,different autologous blood fluids are used during the contacting andincubating steps of a method for preparing a thrombin serum. In anembodiment, a containment device can be a containment device asdescribed herein.

In an embodiment, a method of preparing a thrombin serum can includemultiple incubations and agitations. Following initial contact, theautologous blood fluid and an activator can be mixed (e.g., inversion ofthe containment device, inverting the containment device 5 to 10 times),and in an embodiment, gently mixed. Following mixture, the autologousblood fluid and the activator can be incubated. In an embodiment, theincubation can be about 5 to about 15 minutes, about 5 to about 10minutes, about 5 to about 9 minutes, about 5 to about 8 minutes, about 5to about 7 minutes, about 5 to about 6 minutes, about 6 to about 15minutes, about 6 to about 10 minutes, about 6 to about 9 minutes, about6 to about 8 minutes, or about 6 to about 7 minutes. In an embodiment,an autologous blood fluid and an activator can be incubated for about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, or about 15 minutes following mixture.

Following the first incubation, an additional blood product canoptionally be added to the autologous blood product and activator. Ifthe autologous blood product includes an anticoagulant, then CaC12 isadded following the first incubation. The mixture is then vigorouslymixed (e.g., vortexing, shaking, sharp inversion, etc.). If noadditional blood product or CaC12 is added, the combination ofautologous blood product and activator is vigorously mixed as well.Following the vigorous mixing, the mixture is incubated for about 1,about 2, about 3, about 4, or about 5 minutes. The mixture can then bevigorously mixed again for about 3 to 15 minutes, about 3 to 10 minutes,about 5 to 10 minutes, or about 5 to 15 minutes. In an embodiment, anincubation is about 3, about 4, about 5, about 6, about 7, about 8,about 9, about 10, about 11, about 12, about 13, about 14, or about 15minutes. The mixture can be incubated until at least a clot begins toform. Following a last incubation, thrombin serum is withdrawn from thecontainment device.

In an embodiment of the method to prepare a thrombin serum, all stepsare performed at room temperature.

The exemplary system 10 may include a containment device 12, a cage 14,a cap 16, an inlet port 18 and an outlet port 20. In a non-limitingembodiment, the containment device 12 is configured as a test tube.However, containment devices having other sizes, shapes andconfigurations are also contemplated within the scope of thisdisclosure. In another non-limiting embodiment, the containment device12 is made of a sterilizable material, such as any suitable glass,ceramic or plastic material. In yet another non-limiting embodiment, thecontainment device 12 is made of a transparent material for visualizingthe contents of the containment device 12 during its use.

The containment device 12 extends along a longitudinal axis A between aproximal opening 22 and a closed distal end 24. An internal volume V ofthe containment device 12 is configured to hold an activator. In anembodiment, an activator can be a plurality of beads 30 (see FIG. 1 ).In a non-limiting embodiment, the beads 30 are borosilicate beads, metalbeads, or plastic beads; however, the beads 30 may be manufactured fromany glass like composition including but not limited to alumina,silicate, quartz, bioglass, ceramic glass, flint glass, fluorosilicateglass, phosphosilicate glass, cobalt glass or conundrum. The beads 30may also be spherical shaped to provide for maximum surface area contactwith an autologous blood fluid F that can also be added to the internalvolume V of the containment device 12.

The beads 30 are optionally coated to maximize interaction with theautologous blood fluid F. The coating could be a hydrophilic orhydrophobic surface coating and could include silane, surfactants,polyether, polyester, polyurethane, or polyol groups, for example. Thecoating may optionally be applied to one or more of the beads 30, thecage 14, and an inner surface of the containment device 12.

In another non-limiting embodiment, the beads 30 include a first amountof beads having a first size Si and a second amount of beads having asecond size S2 that is larger than the first size S1. For example, in afurther non-limiting embodiment, the containment device 12 holdsapproximately 1.5 g of 200 um borosilicate beads and approximately 2500mg of 3 mm borosilicate beads. However, other bead amounts and beadsizes are contemplated within the scope of this disclosure, and thespecific bead characteristics may be tailored to match the amount ofdesired interaction between the beads 30 and the autologous blood fluidF received inside the containment device 12.

The cap 16 may be either fixedly or removably attachable to thecontainment device 12 to cover the proximal opening 22 and selectivelyconceal the contents of the containment device 12. In a non-limitingembodiment, the cap 16 is threadably attached to the containment device12. In another non-limiting embodiment, the cap 16 is press-fit onto thecontainment device 12. Other containment device-to-cap connections arealso contemplated within the scope of this disclosure.

The inlet port 18 and the outlet port 20 are received through openings21 formed in the cap 16. In a non-limiting embodiment, the inlet port 18and the outlet port 20 are luer-type connectors adapted for lockinglyengaging a tip of a syringe (syringe not shown in FIGS. 1 and 2 ). Theinlet port 18 may be used to deliver autologous blood fluids F into theinternal volume V of the containment device 12. The outlet port 20 maybe used to remove a thrombin serum from the containment device 12 afterthe autologous blood fluid F has been exposed to and has interacted withthe beads 30 to produce thrombin. The thrombin serum can then beretrieved through the outlet port 20 for subsequent use to create in aclot. The inlet port 18 and the outlet port 20 are swabbable valves, inanother non-limiting embodiment.

Referring now to FIGS. 1-3 , the cage 14 is positioned inside thecontainment device 12. Among other functions, the cage 14 preventsclogging of the inlet port 18 and the outlet port 20. In other words,the cage 14 can act as a filter to prevent clogging. In a non-limitingembodiment, the cage 14 is positioned within a flared portion 26 of thecontainment device 12. The flared portion 26 is proximate to theproximal opening 22. The cage 14 may be either securely affixed (e.g.,welded, etc.) inside the containment device 12 or removable from thecontainment device 12. The cage 14 may include legs 15 that aid toposition and/or secure the cage 14 inside the containment device 12.Slots 17 extend between the legs 15. The cage 14 can include any numberof legs 15 and slots 17.

The cage 14 may further include a cylindrical body 28, although othershapes are also contemplated within the scope of this disclosure. In afirst non-limiting embodiment, the cylindrical body 28 is a hollowcylinder that includes an open top and open bottom (see FIG. 4 ). Thus,in some embodiments, the cage 14 is a floorless structure. A pluralityof openings 32 may extend through the cylindrical body 28 of the cage14. Once introduced into the containment device 12, the autologous bloodfluid F may pass through the openings 32 and be exposed to the beads 30that have already been positioned inside the containment device 12.Exposure to the beads 30 artificially starts the coagulation cascadewithin the autologous blood fluid F. The cells of the autologous bloodfluid F thus release thrombin which can be harvested from the autologousblood fluid F and used to produce a clot, as is further described below.

Referring to FIG. 4 , a filter 35 may optionally attach to either theoutlet port 20 or the opening 21 that receives the outlet port 20. Thefilter 35 can connect to either the outlet port 20 or the opening 21 viaa luer-lock connection, for example. In a further non-limitingembodiment, the filter 35 is a macro-filter that substantially preventsthe beads 30 from clogging the outlet port 20 during extraction of thethrombin serum.

FIGS. 5A and 5B illustrate an optional tray assembly 34 for convenientlypackaging the various components of the system 10. For example, the trayassembly 34 may package the containment device 12, a syringe 36, aneedle 38, and hand warmers 40 of the system 10. In a first non-limitingembodiment, the tray assembly 34 includes a first housing 42 that isconnected to a second housing 46 along a hinge 44. The first housing 42is foldable about the hinge 44 to a position over top of the secondhousing 46 to enclose the system 10. In another non-limiting embodiment,the first housing 42 is separate from and connectable to the secondhousing 46, such as by using a snap-fit or interference connection. Eachhousing 42, 46 includes one or more receptacles 48 for receiving one ormore of the containment device 12, the syringe 36, the needle 38, thehand warmers 40 and/or any other component of the system 10. The firstand second housings 42, 46 may be made of an insulating material.

In another non-limiting embodiment, the tray assembly 34 is employableas a portable incubator. For example, after adding the beads 30 and theautologous blood fluid F to the containment device 12, the hand warmers40 are activated in a known manner and the containment device 12 isplaced inside the tray assembly 34 along with the activated hand warmers40. In a non-limiting embodiment, each hand warmer 40 is positionedwithin one of the receptacles 48 such that it is between the trayassembly 34 and the containment device 12. The tray assembly 34 is thenconcealed by connecting the first housing 42 to the second housing 46.The hand warmers 40 release heat that augments interaction/incubationbetween the autologous blood fluid F and the beads 30, thus promotingthe production of a thrombin serum. The thrombin serum can be extractedfrom the containment device 12 using the syringe 36 and can subsequentlybe used to produce a clot.

FIGS. 6 through 13B, with continued reference to FIGS. 1-5B,schematically illustrate an exemplary surgical technique for preparing athrombin serum and utilizing the thrombin serum to produce a clot. Thesefigures illustrate, in sequential order, a non-limiting embodiment forpreparing a thrombin serum that can be used to produce a clot in anautologous blood fluid. It should be understood; however, that fewer oradditional steps than are recited below could be performed and that therecited order of steps is not intended to limit this disclosure.

Referring first to FIG. 6 , a sample 50 of an autologous blood fluid Fmay be harvested from a body 52 of a patient P. The sample 50 may beharvested from veinous whole blood of the patient P or from a bone ofthe Patient P. In a non-limiting embodiment, the sample 50 is collectedusing the syringe 36 and the needle 38 of the system 10.

The sample 50 can be used to prepare a blood-derived autologous bloodfluid F, such as PRP, PPP, or a combination of these fluids. Forexample, the autologous blood fluid F may be prepared having aconcentrated platelet formulation. Various preparation techniques mayoptionally be performed on the sample 50 to prepare an autologous bloodfluid F having a customized platelet formulation. By way of twonon-limiting examples, the autologous blood fluid F could optionally beprepared using the Arthrex Angel System™ or the Arthrex ACP® System,both available from Arthrex, Inc.

Referring now to FIG. 7 , an autologous blood fluid F is introduced intothe containment device 12. The autologous blood fluid F may includewhole blood, PRP, PPP, or any combinations of these fluids. In anon-limiting embodiment, the autologous blood fluid F that is added tothe containment device 12 is a non-anti-coagulated autologous bloodfluid in that it does not contain any anticoagulants such as AcidCitrate Dextrose Solution A (ACDA). Alternatively, if an autologousblood fluid F having an anticoagulant is used, calcium chloride may beadded to the autologous blood fluid F to overcome the effects of theanticoagulant.

In either case, the autologous blood fluid F may be introduced into thecontainment device 12 by connecting the syringe 36 to the inlet port 18and then injecting the conditioned autologous blood fluid F into theinternal volume V of the containment device 12. The beads 30 and theautologous blood fluid F are exposed to one another inside thecontainment device 12. This exposure artificially starts the coagulationcascade and therefore causes the cells within the autologous blood fluidF to begin to produce thrombin.

The containment device 12 may next be gently inverted and then incubatedfor approximately six to twenty minutes. In a first non-limitingembodiment, the containment device 12 is incubated until a gel isformed, which may occur after approximately seven minutes or longer atambient conditions. The containment device 12 may be incubated on atable top at room temperature. In another non-limiting embodiment, thecontainment device 12 is incubated within an incubation device 60 asschematically shown in FIG. 8 . The incubation device 60 could be anyknown incubator. In yet another non-limiting embodiment, the incubationdevice 60 is the tray assembly 34 of the system 10 (see FIGS. 5A and5B). The tray assembly 34 and the hand warmers 40 provide a portableincubation device. The containment device 12 may be incubated for asuitable amount of time at a suitable temperature to augment theproduction of thrombin within the autologous blood fluid F. Thecontainment device 12 may be incubated at room temperature forapproximately one additional minute to ensure all reactions arecomplete.

An additional amount of the autologous blood fluid F is next added tothe containment device 12 as shown in FIG. 9 . The containment device 12may then be shaken vigorously for approximately ten seconds and thenincubated a second time for approximately one minute (see FIG. 10 ).These amounts of time are considered exemplary and are not intended tolimit this disclosure. The procedure of shaking and incubating may berepeated to ensure that a clot forms in the autologous blood fluid F.The containment device 12 is then gently tapped to break the clot thathas formed inside (schematically shown in FIG. 10 ).

FIG. 11 illustrates removal of a thrombin serum S from the containmentdevice 12. The thrombin serum S can be extracted through the outlet port20 of the containment device 12 by attaching another syringe 55 to theoutlet port 20 and actuating a plunger of the syringe 55.

Referring now to FIGS. 12A and 12B, the thrombin serum S may be mixedwith a second autologous blood fluid F2 in a dish 70. The secondautologous blood fluid F2 may be the same or a different type of fluidas/from the autologous blood fluid F originally added to the containmentdevice 12. In a non-limiting embodiment, the second autologous bloodfluid F2 is a non-anti-coagulated autologous blood fluid. The secondautologous blood fluid F2 could include whole blood, PRP, PPP, or anycombinations of these fluids.

In another non-limiting embodiment, three parts of the second autologousblood fluid F are mixed with one part of the thrombin serum S within thedish (see FIG. 12A). Other ratios are also contemplated, including butnot limited to rations of 1:1, 1:3, 1:4, 1:10, 1:11, etc. After arelatively short period of time, the mixture of the second autologousblood fluid F2 and the thrombin serum S produces a clotted product C(see FIG. 12B). The clotted product C may then be removed from the dish70.

The clotted product C has many potential surgical uses. In a firstnon-limiting embodiment, shown in FIG. 13A, the clotted product C canplaced over top of an implant 80 that has been inserted into tissue 82(e.g., soft or hard tissue including bone) to augment a tissue repair.The clotted product C could optionally be threaded onto a suture 84 toaid in its placement. In another non-limiting embodiment, shown in FIG.13B, the clotted product C can be used to repair or plug a void 86 intissue 82. Other exemplary surgical uses include using the clottedproduct C to treat a wound or to control bleeding.

Although the different non-limiting embodiments are illustrated ashaving specific components, the embodiments of this disclosure are notlimited to those particular combinations. It is possible to use some ofthe components or features from any of the non-limiting embodiments incombination with features or components from any of the othernon-limiting embodiments. Indeed, the embodiments, examples andalternatives of the preceding paragraphs, the claims, or the followingdescription and drawings, including any of their various aspects orrespective individual features, may be practiced independently or in anycombination. Features described in connection with one embodiment areapplicable to all embodiments, unless such features are incompatible.

It should be understood that like reference numerals identifycorresponding or similar elements throughout the several drawings. Itshould also be understood that although a particular componentarrangement is disclosed and illustrated in these exemplary embodiments,other arrangements could also benefit from the teachings of thisdisclosure.

The foregoing description shall be interpreted as illustrative and notin any limiting sense. A worker of ordinary skill in the art wouldunderstand that certain modifications could come within the scope ofthis disclosure. For these reasons, the following claims should bestudied to determine the true scope and content of this disclosure.

The invention claimed is:
 1. A method for preparing a clotted bloodproduct, comprising: incubating a first autologous blood fluid in acontainment device, wherein the containment device comprises: (a) alongitudinal axis; (b) a cage received within said containment device,the cage having a wall or walls that extend along a longitudinal axisinto the containment device, wherein the cage wall or walls include aplurality of openings; (c) a cap attached to said containment device;(d) an inlet port configured to introduce the first autologous bloodfluid into said containment device, wherein the inlet port is locatedradially outwardly of the cage relative to the longitudinal axis, suchthat a fluid is delivered to the containment device between an innerwall of the containment device and an outer wall of the cage; and (e) anoutlet port configured to remove a thrombin serum from said containmentdevice; extracting a thrombin serum from the containment device; addingthe thrombin serum to a second autologous blood fluid to produce aclotted product; and collecting the clotted product.
 2. The method ofclaim 1, wherein the clotted product is placed over the top of animplant in a patient.
 3. The method of claim 1, wherein the clottedproduct is threaded onto a suture.
 4. The method of claim 1, wherein theclotted product is placed into a void in a tissue of a patient.
 5. Themethod of claim 1, wherein the first autologous blood fluid is anon-anticoagulated autologous blood fluid.
 6. The method of claim 1,wherein the second autologous blood fluid is a non-anticoagulatedautologous blood fluid.
 7. The method of claim 1, wherein the firstautologous blood fluid is whole blood, platelet rich plasma, plateletpoor plasma, or a combination thereof.
 8. The method of claim 1, whereinthe second autologous blood fluid is whole blood, platelet rich plasma,platelet poor plasma, or a combination thereof.
 9. The method of claim1, comprising, prior to extracting the thrombin serum, adding anadditional amount of the first autologous blood fluid to the containmentdevice; shaking or inverting the containment device; and incubating thecontainment device a second time.
 10. The method of claim 9, furthercomprising shaking or inverting the containment device a second time,and incubating the containment device a third time.
 11. The method ofclaim 1, comprising, prior to adding the first autologous blood fluid,adding a plurality of beads to the containment device.
 12. The method ofclaim 11, wherein the plurality of beads are borosilicate beads, metalbeads, or plastic beads.
 13. The method of claim 11, comprising ahydrophilic or hydrophobic surface coating on at least one of theplurality of beads, the cage, or an inner surface of the containmentdevice.
 14. The method of claim 1, wherein the second autologous bloodfluid is a different type of fluid than the first autologous bloodfluid.
 15. The method of claim 1, wherein the outlet port comprises afilter attached to either the outlet port or to an opening that receivesthe outlet port.